EP0446826A2 - ProcédÀ© pour la préparation enzymatique de cyannydrine optiquement active - Google Patents

ProcédÀ© pour la préparation enzymatique de cyannydrine optiquement active Download PDF

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Publication number
EP0446826A2
EP0446826A2 EP91103667A EP91103667A EP0446826A2 EP 0446826 A2 EP0446826 A2 EP 0446826A2 EP 91103667 A EP91103667 A EP 91103667A EP 91103667 A EP91103667 A EP 91103667A EP 0446826 A2 EP0446826 A2 EP 0446826A2
Authority
EP
European Patent Office
Prior art keywords
liquid crystal
reaction
reactor
liquid
organic solvent
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
EP91103667A
Other languages
German (de)
English (en)
Other versions
EP0446826B1 (fr
EP0446826A3 (en
Inventor
Peter Dr. Miethe
Maria Regina Prof. Kula
Ingeborg Maria Stürtz
Christian Prof. Dr. Wandrey
Udo Kragl
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Forschungszentrum Juelich GmbH
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Forschungszentrum Juelich GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from DE19904008412 external-priority patent/DE4008412C1/de
Priority claimed from DE19904008411 external-priority patent/DE4008411A1/de
Priority claimed from DE19904028689 external-priority patent/DE4028689A1/de
Application filed by Forschungszentrum Juelich GmbH filed Critical Forschungszentrum Juelich GmbH
Publication of EP0446826A2 publication Critical patent/EP0446826A2/fr
Publication of EP0446826A3 publication Critical patent/EP0446826A3/de
Application granted granted Critical
Publication of EP0446826B1 publication Critical patent/EP0446826B1/fr
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Expired - Lifetime legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/002—Nitriles (-CN)
    • C12P13/004—Cyanohydrins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/18—Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2219/00—Aspects relating to the form of the liquid crystal [LC] material, or by the technical area in which LC material are used
    • C09K2219/15—Aspects relating to the form of the liquid crystal [LC] material, or by the technical area in which LC material are used used as a medium, in which chemical reactions take place

Definitions

  • the invention relates to a process for the production of optically active cyanohydrins by enzymatic reaction of oxone compounds with hydrocyanic acid in the presence of (R) - or (S) -oxynitrilase (4.1.2.10) or (4.1.2.11) under such acidic conditions that the chemical competition and racemization are negligible.
  • Optically active cyanohydrins are of considerable interest for the production of optically active ⁇ -amino alcohols, ⁇ -hydroxycarboxylic acids, heterocycles and pyrethroid insecticides.
  • DE-PS 13 00 111 describes the production of optically active cyanohydrins with (R) -oxynitrilase bound to ion exchangers at pH 5.4 .
  • the ee values achieved were consistently below 90%.
  • Effenberger et al (Angew. Chem. 99 (1987) 491) therefore recommends the enzymatic reaction of oxo compounds with hydrocyanic acid in organic water-immiscible solvents in order to suppress the chemical reaction. It was preferred to work with carrier-fixed enzyme in ethyl acetate at pH 5.4. Ee values of up to 99% were achieved.
  • the process according to the invention of the type mentioned at the outset is therefore characterized in that the reaction is carried out in an organic solvent in the presence of oxynitrilase solubilized in a lyotropic liquid crystal, surfactants whose hydrolysis leading to an increase in the pH being excluded for liquid crystal formation .
  • optically active (R) -cyanohydrins and (S) -cyanohydrins can be obtained by reacting aliphatic, aromatic or heteroaromatic aldehydes or ketones with hydrocyanic acid in the presence of the (R) - or (S) -oxynitrilase "immobilized" in the lyotropic liquid crystal be preserved.
  • buffer solution for the production of the liquid crystalline phase, in particular buffer solution is used, the pH of which is between 3 and 6.
  • Suitable surfactants of cationic or nonionic amphiphiles are preferred, such as alkyltrimethylammonium halides, alkyl pyridinium halides, polyoxyethylene ethers, polyoxyethylene esters, polyoxyethylene sorbitan esters, alkylphenol polyethylene glycol ethers, corresponding polyoxypropylene derivatives or Copolyoxyethylenpolyoxypropylenderivate or corresponding mixtures of surfactants and organic solvents are especially methylene chloride, chloroform, carbon tetrachloride, dibutyl ether, diisopropyl ether or toluene appropriate.
  • dibutyl ether and Brij® 35 polyoxyethylene monolauryl ether
  • liquid-crystalline biocatalyst-containing system it is expedient to start with a 1 to 30% by weight, preferably 5 to 10% by weight, stock solution or suspension of a Surfactant in an organic solvent. 1 to 30% by weight, in particular 5 to 10% by weight, of aqueous biocatalyst-containing buffer solution are subsequently added to this stock solution. After a brief shake, the biphase liquid crystal / organic solvent system is formed.
  • composition of such a system must be determined in corresponding preliminary tests, unless it can be found in the colloid chemical literature (ternary phase diagrams surfactant / organic solvent / water).
  • the invention is particularly suitable for the implementation of water-insoluble oxo compounds. If the oxo compound is not too polar, it can itself form the organic solvent.
  • biocatalyst-containing lyotropic liquid crystals are preferably mixed with porous support materials, such as. B. porous glass sintered bodies, the use in the form of a packed column is particularly useful.
  • a continuous-flow reactor which has thin layers of the biocatalyst-containing liquid crystal adjacent to narrow flow channels for substrate-containing solvent is also particularly advantageous in terms of process technology.
  • optical purity 99% ee The optical purity was determined as N, O-bis (pentafluoropropionyl) -2-amino-1-phenylethanol derivative of the R-mandelonitrile by capillary gas chromatography according to H. Frank et al. (J. Chromatogr. 146, (1987), 197-206) on a chiral separation phase (FS-Chirasil-Val, 25 mx 0.32 mm).
  • the derivatization was carried out as follows: 1-2 mg of the mandelonitrile were reduced with 250 ⁇ l of a 1 M diborane solution (in tetrahydrofuran) in dibutyl ether at room temperature in 30 minutes. After hydrolysis of the excess diborane with a few drops of ethanol and removal of the solvent, the amino alcohol obtained was acylated directly with 20 ⁇ l of pentafluoropropionic anhydride in methylene chloride at room temperature in 15 minutes. Finally, excess anhydride was removed on a rotary evaporator, the residue was taken up again with methylene chloride and analyzed by gas chromatography.
  • Example 1 100 mg of 3-phenoxybenzaldehyde were reacted. The reaction was complete after about 90 minutes. The cyanohydrin was worked up and the optical purity was determined as described in Example 1. Chemical yield 90 mg (87% of theory) Optical purity 99% ee
  • Example 1 100 mg of 3-fluorobenzaldehyde were reacted. The reaction was complete after 90 minutes. Working up was carried out analogously to Example 1. Chemical yield 102 mg (83% of theory) Optical purity 76% ee
  • Example 1 100 mg of 2-methylcyclohexanone were reacted. The reaction was complete after about 120 minutes. Working up and analysis were carried out analogously to Example 1. Chemical yield 111 mg (82% of theory) Optical purity 92% de
  • a solution of 25% by weight of Brij 35 (SIGMA) in dibutyl ether was prepared.
  • 3.5 ml of an aqueous (R) -oxynitrilase solution were added to 10 ml of this solution (protein concentration 10 mg / ml, citrate buffer pH 3.75).
  • R aqueous
  • lyotropic liquid crystal are formed in equilibrium with dibutyl ether.
  • the liquid-crystalline biocatalyst was mixed with 70 g of glass beads and placed in a 50 ml chromatography column.
  • the column effluent was connected to a pump followed by a bubble trap and polarimetric flow cell, and the system was filled with 50 ml of dibutyl ether.
  • the reaction in the flow apparatus was started by switching on the pump. Different dwell times were set and each waited until steady state was reached. The apparatus was operated continuously for 3 days, with hardly any drop in the rotational value.
  • the volume that can be flowed through was used to calculate the residence time and the space-time yield.
  • the conversion and the rotation value were determined at 10 different residence times.
  • a reactor for such reactions using biocatalysts which are used in the form of lyotropic liquid crystals e.g. a flow reactor expedient in which alternate layers containing liquid crystals alternate with flow channels for substrate-containing liquid, which are at least partially formed by liquid-permeable layers made of porous material which delimits the liquid crystal-containing layers, the dimensions of which transversely to the flow channels result in a substantially complete penetration of the entire liquid crystal-containing layer allow reactive substrate.
  • the flow channels are formed in particular by layers of liquid-permeable sintered material and preferably fill ⁇ 50%, in particular ⁇ 30% of the reactor volume.
  • the liquid crystal-containing layer is formed in particular by a mixture of liquid crystals containing biocatalyst with porous carrier material, in particular porous sintered glass, and preferably has a layer thickness of ⁇ 1 cm. That of the flow channels is preferably ⁇ 0.5 cm.
  • FIG. 1 shows a reactor 1, the porous tube 2 of which contains a liquid crystal filling 3 having a biocatalyst and is closed at the end by caps, plugs or the like 4, 5.
  • the tube 2 slidably fits into the cladding tube 6, which forms the reactor jacket and forms a flow channel for liquid flow, which is formed by the gap 7 together with the pore volume of the tube 2.
  • the liquid flows in via an inlet distributor 8 and exits the reactor via 9. End closure and liquid distributor are not specified.
  • FIG. 2 shows the structure of a plate module reactor 10, which is formed by a series of plates 20 which can be flowed through in the same direction or alternating (as sketched) and between which liquid crystal layers 30 are provided.
  • the simplest of such a reactor consisting of a plurality of plates is formed by a stack of mutually independent plates 20, the longitudinal edges of which are sealed off with the reactor wall 60.
  • Liquid crystal layers are applied to the large surfaces 20 'of the plates 20 on one or both sides, the thickness of which determines the space between the plates, with spacers, if necessary, ensuring that the selected distance is maintained.

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  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Sustainable Development (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
EP91103667A 1990-03-16 1991-03-11 Procédé pour la préparation enzymatique de cyannydrine optiquement active Expired - Lifetime EP0446826B1 (fr)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
DE4008412 1990-03-16
DE19904008412 DE4008412C1 (en) 1990-03-16 1990-03-16 Optically active cyanohydrin prepn. useful insecticidal derivs. - comprising condensation of oxo-cpd. with hydrogen cyanide in presence of (R)- or (S) oxynitrilase
DE4008411 1990-03-16
DE19904008411 DE4008411A1 (de) 1990-03-16 1990-03-16 Durchflussreaktor fuer biokatalytische umsetzungen in gegenwart lyotroper fluessigkristalle
DE4028689 1990-09-10
DE19904028689 DE4028689A1 (de) 1990-09-10 1990-09-10 Verfahren zur enzymatischen herstellung optisch aktiver cyanhydrine

Publications (3)

Publication Number Publication Date
EP0446826A2 true EP0446826A2 (fr) 1991-09-18
EP0446826A3 EP0446826A3 (en) 1992-07-15
EP0446826B1 EP0446826B1 (fr) 1995-11-15

Family

ID=27200987

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EP91103667A Expired - Lifetime EP0446826B1 (fr) 1990-03-16 1991-03-11 Procédé pour la préparation enzymatique de cyannydrine optiquement active

Country Status (5)

Country Link
US (1) US5122462A (fr)
EP (1) EP0446826B1 (fr)
JP (1) JPH0638793A (fr)
DE (1) DE59106876D1 (fr)
DK (1) DK0446826T3 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0547655A1 (fr) * 1991-12-11 1993-06-23 Duphar International Research B.V Méthode de préparation de cyanohydrines optiquement actives
DE19529116A1 (de) * 1995-08-08 1997-03-06 Chemie Linz Deutschland Gmbh I (S)-Hydroxynitrillyase aus Hevea brasiliensis
EP0799894A3 (fr) * 1996-02-09 1999-12-08 Degussa-Hüls Aktiengesellschaft Procédé de préparation de (S)-cyanhydrines
WO2005040393A1 (fr) * 2003-10-10 2005-05-06 Degussa Ag Procede de preparation d'acides alpha-carboxyliques et d'amides enrichis en enantiomeres

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AT396252B (de) * 1991-10-31 1993-07-26 Chemie Linz Gmbh Enzymatisches verfahren zur enantioselektiven herstellung optisch aktiver cyanhydrine
US5177242A (en) * 1991-12-17 1993-01-05 Fmc Corporation Process for preparing optically active cyanohydrins with enzymes
US5241087A (en) * 1992-03-09 1993-08-31 Bend Research, Inc. Enantiomeric enrichment of cyanohydrins
AT400035B (de) * 1993-06-01 1995-09-25 Chemie Linz Gmbh Enzymatisches verfahren zur herstellung aliphatischer s-cyanhydrine
JP4497081B2 (ja) 2005-10-31 2010-07-07 トヨタ自動車株式会社 人の状態検出装置
CN101827810A (zh) * 2007-10-23 2010-09-08 日宝化学株式会社 羟腈化合物的制备方法与制备装置,以及α-羟基酯化合物的制备方法

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1300111B (de) * 1965-04-07 1969-07-31 Pfeil Verfahren zur Herstellung von optisch aktiven Cyanhydrinen
DE3222912A1 (de) * 1982-06-18 1983-12-22 Basf Ag, 6700 Ludwigshafen Unloeslicher biokatalysator
DE3701383A1 (de) * 1987-01-20 1988-07-28 Degussa Verfahren zur herstellung von optisch aktiven cyanhydrinen
US4956289A (en) * 1987-03-16 1990-09-11 Brunswick Corporation Thin film membrane enzyme reactor and method of using same
US4800162A (en) * 1987-04-01 1989-01-24 Sepracor, Inc. Method for resolution of steroisomers in multiphase and extractive membrane reactors
DE3823864A1 (de) * 1988-01-29 1989-08-10 Kernforschungsanlage Juelich Enymatisches verfahren zur herstellung von optisch aktiven cyanhydrinen
DD282822A7 (de) * 1988-05-06 1990-09-26 Univ Halle Wittenberg Verfahren zur biokatalytischen umsetzung schlecht wasserloeslicher substanzen
DD278478A3 (de) * 1988-06-23 1990-05-09 Univ Halle Wittenberg Verfahren zur biokatalytischen herstellung schlecht wasserloeslicher substanzen mit integrierter extraktiver aufarbeitung
DE3917374A1 (de) * 1988-07-14 1990-12-06 Forschungszentrum Juelich Gmbh Verfahren zur herstellung von s-cyanhydrinen

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0547655A1 (fr) * 1991-12-11 1993-06-23 Duphar International Research B.V Méthode de préparation de cyanohydrines optiquement actives
DE19529116A1 (de) * 1995-08-08 1997-03-06 Chemie Linz Deutschland Gmbh I (S)-Hydroxynitrillyase aus Hevea brasiliensis
EP0799894A3 (fr) * 1996-02-09 1999-12-08 Degussa-Hüls Aktiengesellschaft Procédé de préparation de (S)-cyanhydrines
WO2005040393A1 (fr) * 2003-10-10 2005-05-06 Degussa Ag Procede de preparation d'acides alpha-carboxyliques et d'amides enrichis en enantiomeres

Also Published As

Publication number Publication date
US5122462A (en) 1992-06-16
DK0446826T3 (da) 1996-02-12
EP0446826B1 (fr) 1995-11-15
JPH0638793A (ja) 1994-02-15
EP0446826A3 (en) 1992-07-15
DE59106876D1 (de) 1995-12-21

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